The minimum detectable dose of this kit is typically less than 33pg/mL.

Specificity

This assay has high sensitivity and excellent specificity for detection of Vinculin (VCL).
No significant cross-reactivity or interference between Vinculin (VCL) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Vinculin (VCL) and the recovery rates were calculated by comparing the measured value to the expected amount of Vinculin (VCL) in samples.

Matrix

Recovery range (%)

Average(%)

serum(n=5)

78-98

93

EDTA plasma(n=5)

78-88

83

heparin plasma(n=5)

95-102

99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Vinculin (VCL) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Vinculin (VCL) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Vinculin (VCL) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample

1:2

1:4

1:8

1:16

serum(n=5)

93-101%

80-91%

91-98%

83-96%

EDTA plasma(n=5)

85-104%

86-95%

95-102%

86-97%

heparin plasma(n=5)

83-99%

84-104%

80-102%

92-105%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents

Quantity

Reagents

Quantity

Pre-coated, ready to use 96-well strip plate

1

Plate sealer for 96 wells

4

Standard

2

Standard Diluent

1×20mL

Detection Reagent A

1×120µL

Assay Diluent A

1×12mL

Detection Reagent B

1×120µL

Assay Diluent B

1×12mL

TMB Substrate

1×9mL

Stop Solution

1×6mL

Wash Buffer (30 × concentrate)

1×20mL

Instruction manual

1

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Vinculin (VCL). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Vinculin (VCL). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Vinculin (VCL), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Vinculin (VCL) in the samples is then determined by comparing the O.D. of the samples to the standard curve.